人骨髓间充质干细胞向汗腺细胞诱导培养中基因表达方式的研究

Gene expression in culture to induce human bone marrow mesenchymal stem cells into sweat glands cells

目的通过诱导人骨髓间充质干细胞(hMSCs)向汗腺细胞(hSGCs)分化并检测经诱导后细胞的基因表达变化,探讨hMSCs的可塑性分化机制。方法分离纯化并体外扩增hMSCs,设立对照组(DM组)和向hSGCs诱导的SG组、EGF10组和EGF20组等四组不同培养基体系,观测细胞形态学变化和采用流式细胞仪、RT-PCR等方法检测融合细胞的基因和蛋白表达特征。结果细胞在不同条件培养液中诱导培养均能存活;诱导培养2d后细胞数量开始扩增,细胞形态也由间充质干细胞的长梭状开始向hSGCs的扁平状转变,在SG组尤其明显;诱导培养1周后细胞数量扩增明显;与汗腺发生形成密切的基因EGF和EGFR出现高丰度表达,并且表达了仅hSGCs才有表达的CK7蛋白。结论 hMSCs在不同介质诱导培养下可在形态学和分子水平上呈现向hSGCs横向分化的潜能,hMSCs的可塑性现象丰富了皮肤创面修复和汗腺再生理论。

Objective To study the underlying plasticity mechanism of human bone marrow mesenchymal stem cells（hMSC） by analyzing the differentiation of hMSC into sweat glands cells（hSGC） and detecting the changes in gene expression.Methods The HMSC were divided into control group（DM group）,hSGC-induced SG group,EGF10 group and EGF20 group after they were isolated,purified and amplified.Changes in morphology of HMSC were observed,and expressions of genes and proteins in fused HMSC were detected by flow cytometry and RT-PCR,respectively.Results The HMSC could survive in different cultured fluids.The number of HMSC was increased 2d after induced culture,and the long spindle-like HMSC changed into flat hSGC,which was especially significant in SG group.The number of HMSC was significantly increased one week after induced culture.EGF and EGFR,closely related with sweat glands,were significantly expressed in hMSC,and CK7 protein was only expressed in hSGC.Conclusion HMSC,cultured with different media,can differentiate into hSGC at morphologic and molecular levels.The plasticity of hMSC may enrich the theory in regulating repair and rebuilding of skin and sweat glands.